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Journal of Lipid Research, Vol 26, 1324-1332, Copyright © 1985 by Lipid Research, Inc.

ARTICLES

Bile acid synthesis in rat liver peroxisomes: metabolism of 26- hydroxycholesterol to 3 beta-hydroxy-5-cholenoic acid

SK Krisans, SL Thompson, LA Pena, E Kok and NB Javitt

Rat liver peroxisomes have been found to oxidize 26-hydroxycholesterol, the product of cholesterol C-26 hydroxylation to 3 beta-hydroxy-5- cholenoic acid. Peroxisomes were purified by differential and equilibrium density centrifugation in a steep linear metrizamide gradient to greater than 95% purity. Purity of peroxisomes was determined by measurement of specific marker enzymes. The activities of cytochrome oxidase (a mitochondrial marker) and acid phosphatase (a lysosomal marker) in the purified peroxisome fractions were below the level of detection. Esterase activity indicated a 2-4% microsomal contamination. Subsequent to incubation of peroxisomes with [16,22-3H]- 26-hydroxycholesterol, the reaction products were extracted, methylated, acetylated, and subjected to thin-layer, high pressure liquid, and gas-liquid chromatographic analyses. 3 beta-Hydroxy-5- cholenoic acid was the major identifiable metabolite of 26- hydroxycholesterol. Incubations of pure microsomal fractions (greater than 99%) with 26-hydroxycholesterol under the same conditions demonstrated that the production of 3 beta-hydroxy-5-cholenoic acid by peroxisomes was not attributable to microsomal contamination. This study demonstrates that peroxisomes participate in the side-chain oxidation of intermediates in bile acid synthesis.
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